Factorial screening of antibody purification processes using three chromatography steps without protein A
Factorial screening of antibody purification processes using three chromatography steps without protein A
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DOI：
10.1016/j.chroma.2003.10.060
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发表时间：
2004-01-23
影响因子：
4.1
通讯作者：
Farner, RL
中科院分区：
文献类型：
--
作者：
Follman, DK;Farner, RL
蛋白A亲和层析通常被用作捕获步骤，以满足药物抗体纯化的纯度、产量和吞吐量要求。然而，在步进性能和价格之间存在权衡。蛋白A树脂可去除99.9%的饲料杂质，但其价格明显高于非亲和介质。由于许多抗体的治疗适应症需要高剂量和/或长期给药，因此过程经济学的考虑是至关重要的。我们系统地评价了阳离子交换、阴离子交换、羟基磷灰石、疏水相互作用、疏水电荷诱导和小分子配基树脂在CHO单抗三步层析纯化过程中的每一步的纯化性能。通过阳离子交换-阴离子交换-疏水作用层析、阳离子交换-阴离子交换-混合阳离子交换层析和阳离子交换-混合阳离子交换-阴离子交换层析，将宿主细胞蛋白质去除到检测不到的程度。工艺步骤的先后顺序对净化效果有显著影响。(C)2003爱思唯尔B.V.保留所有权利。
Protein A affinity chromatography is often employed as a capture step to meet the purity, yield, and throughput requirements for pharmaceutical antibody purification. However, a trade-off exists between step performance and price. Protein A resin removes 99.9% of feed stream impurities; however, its price is significantly greater than those of non-affinity media. With many therapeutic indications for antibodies requiring high doses and/or chronic administration, the consideration of process economics is critical. We have systematically evaluated the purification performance of cation-exchange, anion-exchange, hydroxyapatite, hydrophobic interaction, hydrophobic charge induction, and small-molecule ligand resins in each step of a three-step chromatographic purification process for a CHO-derived monoclonal antibody. Host cell proteins were removed to less-than-detectable for three processes (cation-exchange-anion-exchange-hydrophobic interaction chromatography, cation-exchange-anion-exchange-mixed cation-exchange chromatography, and cation-exchange-mixed cation-exchange-anion-exchange chromatography). The order of the process steps affected purification performance significantly. (C) 2003 Elsevier B.V. All rights reserved.